10.1021/nn401661d.s002
Jung-uk Shim
Jung-uk
Shim
Rohan T. Ranasinghe
Rohan T.
Ranasinghe
Clive A. Smith
Clive A.
Smith
Shehu M. Ibrahim
Shehu M.
Ibrahim
Florian Hollfelder
Florian
Hollfelder
Wilhelm T. S. Huck
Wilhelm
T. S. Huck
David Klenerman
David
Klenerman
Chris Abell
Chris
Abell
Ultrarapid Generation of Femtoliter Microfluidic Droplets for Single-Molecule-Counting Immunoassays
American Chemical Society
2016
analyte molecules
2 orders
prostate cancer
Ultrarapid Generation
10 min
femtoliter droplets
enzyme molecule
Femtoliter Microfluidic Droplets
enzyme molecules
reporter enzyme
chemical reactions
46 fM
microfluidic device
prototype system
biomolecular complexes
2016-02-19 02:59:36
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Ultrarapid_Generation_of_Femtoliter_Microfluidic_Droplets_for_Single_Molecule_Counting_Immunoassays/2393923
We report a microfluidic droplet-based approach enabling the measurement of chemical reactions of individual enzyme molecules and its application to a single-molecule-counting immunoassay. A microfluidic device is used to generate and manipulate <10 fL droplets at rates of up to 1.3 × 10<sup>6</sup> per second, about 2 orders of magnitude faster than has previously been reported. The femtodroplets produced with this device can be used to encapsulate single biomolecular complexes tagged with a reporter enzyme; their small volume enables the fluorescent product of a single enzyme molecule to be detected within 10 min of on-chip incubation. Our prototype system is validated by detection of a biomarker for prostate cancer in buffer, down to a concentration of 46 fM. This work demonstrates a highly flexible and sensitive diagnostic platform that exploits extremely high-speed generation of monodisperse femtoliter droplets for the counting of individual analyte molecules.